1 reply to this topic

[wangml]

Hi, everyone;

I have done a series of PCR and found that there is slight difference between control and treated group according to bands stained with EB in 2% agarose gel. I was told that such a result is much less sensitive than that of southern. So I blotted such gel  to membrane for further southern analysis. Here is the question, if I can use the same PCR product to construct my probe? Yes or No? and How?
Any advice will be appreciated in advance.

[milanoj]

So you want to quantitate the PCR product? No need to southernize the DNA. A great method is Pico Green Quanitiation of your PCR products. It will distinguish nicely between unincorporated primers and dNTPs. See Milano et al. Nucleic Acids Research.